title: Establishment of a soaking RNA interference and Bombyx mori nucleopolyhedrovirus (BmNPV)-hypersensitive cell line using Bme21 cell.
abstract: The double-stranded RNA (dsRNA) mediated RNA interference (RNAi) is widely employed in silkworm and its tissue-derived cell lines for gene function analysis. Baculovirus expression vector system (BEVS) has an advantage for large-scale protein expression. Previously, combining these useful tools, we improved traditional AcMNPV-Sf9 BEVS to produce modified target glycoproteins, where the ectopic expression of Caenorhabditis elegans systemic RNAi defective-1 (SID-1) was found to be valuable for soaking RNAi. In current study, we applied CeSID-1 protein to a Bombyx mori NPV (BmNPV)-hypersensitive Bme21 cell line and investigated its properties both in soaking RNAi ability and recombinant protein expression. The soaking RNAi-mediated suppression in the Bme21 cell enables us to produce modified glycoproteins of interest in BmNPV-Bme21 BEVS.
authors.affiliation: Laboratory of Silkworm Science, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, 6-10-1 Hakozaki Higashi-ku, Fukuoka, 812-8581, Japan.
authors.forename: Jian
authors.initials: J
authors.lastname: Xu
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Mon
authors.forename: Takahiro
authors.initials: T
authors.lastname: Kusakabe
authors.forename: Zhiqing
authors.initials: Z
authors.lastname: Li
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Iiyama
authors.forename: Atsushi
authors.initials: A
authors.lastname: Masuda
authors.forename: Takumi
authors.initials: T
authors.lastname: Mitsudome
authors.forename: Jae Man
authors.initials: JM
authors.lastname: Lee
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D017173:Caenorhabditis elegans / Q000235:genetics; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D018929:Cell Culture Techniques; D006023:Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008967:Molecular Biology / Q000379:methods*; D008969:Molecular Sequence Data; D017924:Nucleopolyhedroviruses / Q000235:genetics; D034622:RNA Interference; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D061987:Sf9 Cells; D018411:Spodoptera
chemical_list: D029742:Caenorhabditis elegans Proteins; D006023:Glycoproteins; D008565:Membrane Proteins; D011994:Recombinant Proteins; C452203:SID-1 protein, C elegans